Impaired retinal blood flow (RBF) autoregulation plays a key role in the development and progression of several ocular diseases, including glaucoma and diabetic retinopathy. Clinically, reproducible RBF quantitation could significantly improve early diagnosis and disease management. Several non-invasive techniques have been developed but are limited for retinal microvasculature flow measurements due to their low signal-to-noise ratio and poor lateral resolution. In this study, we demonstrate reproducible vessel caliber and retinal blood flow velocity measurements in healthy human volunteers using a high-resolution (spatial and temporal) multimodal adaptive optics system with scanning laser ophthalmoscopy and optical coherence tomography.